4.8. Co-Immunoprecipitation

We studied the effect of various treatments as indicated on CBP/β-catenin and p300/β-catenin association by co-immunoprecipitation as previously described [25]. Briefly, pancreatic cancer cells were treated with ICG-001 for 24 h, and cytoplasmic and nuclear protein fractions were extracted. From the nuclear fractions, we immunoprecipitated CBP and p300 on agarose beads. The eluents were run on standard sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to a polyvinylidene difluoride (PVDF) membrane. Membranes were immunoblotted for β-catenin.